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Steinhäuser, Christine; Heigl, Ulrike; Tchikov, Vladimir; Schwarz, Jeanette; Gutsmann, Thomas; Seeger, Katrin; Brandenburg, Julius; Fritsch, Jürgen; Schroeder, Josef; Wiesmüller, Karl‐Heinz; Rosenkrands, Ida; Walther, Paul; Pott, Johanna; Krause, Eberhard; Ehlers, Stefan; Schneider‐Brachert, Wulf; Schütze, Stefan; Reiling, Norbert

Lipid‐Labeling Facilitates a Novel Magnetic Isolation Procedure to Characterize Pathogen‐Containing Phagosomes

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  • Media type: E-Article
  • Title: Lipid‐Labeling Facilitates a Novel Magnetic Isolation Procedure to Characterize Pathogen‐Containing Phagosomes
  • Contributor: Steinhäuser, Christine; Heigl, Ulrike; Tchikov, Vladimir; Schwarz, Jeanette; Gutsmann, Thomas; Seeger, Katrin; Brandenburg, Julius; Fritsch, Jürgen; Schroeder, Josef; Wiesmüller, Karl‐Heinz; Rosenkrands, Ida; Walther, Paul; Pott, Johanna; Krause, Eberhard; Ehlers, Stefan; Schneider‐Brachert, Wulf; Schütze, Stefan; Reiling, Norbert
  • imprint: Wiley, 2013
  • Published in: Traffic
  • Language: English
  • DOI: 10.1111/tra.12031
  • ISSN: 1398-9219; 1600-0854
  • Keywords: Cell Biology ; Genetics ; Molecular Biology ; Biochemistry ; Structural Biology
  • Origination:
  • Footnote:
  • Description: <jats:p><jats:bold>Here we describe a novel approach for the isolation and biochemical characterization of pathogen‐containing compartments from primary cells: We developed a lipid‐based procedure to magnetically label the surface of bacteria and visualized the label by scanning and transmission electron microscopy (<jats:styled-content style="fixed-case">SEM</jats:styled-content>, <jats:styled-content style="fixed-case">TEM</jats:styled-content>). We performed infection experiments with magnetically labeled <jats:italic>Mycobacterium avium</jats:italic>, <jats:italic>M. tuberculosis</jats:italic> and <jats:italic>Listeria monocytogenes</jats:italic> and isolated magnetic bacteria‐containing phagosomes using a strong magnetic field in a novel free‐flow system. Magnetic labeling of <jats:italic>M. tuberculosis</jats:italic> did not affect the virulence characteristics of the bacteria during infection experiments addressing host cell activation, phagosome maturation delay and replication in macrophages <jats:italic>in vitro</jats:italic>. Biochemical analyses of the magnetic phagosome‐containing fractions provided evidence of an enhanced presence of bacterial antigens and a differential distribution of proteins involved in the endocytic pathway over time as well as cytokine‐dependent changes in the phagosomal protein composition. The newly developed method represents a useful approach to characterize and compare pathogen‐containing compartments, in order to identify microbial and host cell targets for novel anti‐infective strategies.</jats:bold></jats:p>
  • Access State: Open Access